LOCUS       sequence-364918-        7014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for ATP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  AUTHORS   Ino D, Tanaka Y, Hibino H, Nishiyama M
  TITLE     A fluorescent sensor for real-time measurement of extracellular 
            oxytocin dynamics in the brain.
  JOURNAL   Nat Methods. 2022 Sep 22. pii: 10.1038/s41592-022-01597-x. doi: 
            10.1038/s41592-022-01597-x.
  PUBMED    36138174
REFERENCE   2  (bases 1 to 7014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2022 Sep 22. pii: 10.1038/s41592-022-01597-x. doi: 
            10.1038/s41592-022-01597-x."
COMMENT     Sequence Label: pCIS.MTRIA-ATP
FEATURES             Location/Qualifiers
     source          1..7014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..581
                     /label=chicken beta-actin promoter
     intron          582..1598
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1522..1544
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1606..1625
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1654..1670
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1655..1671
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2600..2621
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2727..2748)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3541..3557)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3541..3557)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3646..3665)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3711..3766
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3765..3784)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4121..4139)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4160..4176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4160..4176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4173..4195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4184..4200
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4208..4214,4215..4232,4233..4238))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4253..4274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4391..4408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4562..5150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4642..4661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5321..6112,6113..6181))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5944..5963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6182..6286)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6313..6768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6400..6419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6610..6631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6894..6916
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6909..6925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6932..6951
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6932..6950
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      421 gggggsgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
      481 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      541 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      601 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      661 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
      721 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
      781 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
      841 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
      901 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
      961 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     1021 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     1081 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     1141 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     1201 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     1261 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     1321 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     1381 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     1441 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     1501 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     1561 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1621 ttgtgctgtc tcatcatttt ggcaaaggat tcctcgaggt cgacggtatc gataagcttg
     1681 atatcgaatt caccatggca gcagacctgg aaccctggaa tagcaccatc aatggcacct
     1741 gggaggggga cgaactggga tacaagtgtc gcttcaacga ggacttcaag tacgtgctgt
     1801 tgcccgtgtc ctatggcgtg gtgtgcgtgc tcgggttgtg cctgaacgtc gtggctctct
     1861 atatcttcct atgccgcctc aaaacctgga acgcctccac cacctacatg tttcacctgg
     1921 cagtttcgga ctctctctac gcagcgtccc tgccgctgtt ggtttattac tacgcccggg
     1981 gtgaccactg gccatttagc acggtgctct gcaagctggt gcgtttcctc ttctacacca
     2041 acctctactg cagcatcctc ttcctcacct gcatcagcgt gcaccggtgc ctgggagtcc
     2101 tgcgccctct gcactccctg cgttggggcc gggcccgtta tgcccgccgg gtggctgcgg
     2161 ttgtgtgggt gctggtgctg gcctgccagg cacccgtgct ctacttcgtc accaccagcg
     2221 tgcggggaac ccggatcact tgccatgaca cctcggcccg agagctcttt agccattttg
     2281 tggcttacag ctccgtcatg ctgggtctgc tttttgctgt gcccttttcc gtaatcctgg
     2341 tctgttacgt gcttatgtgc tacaaaatat ggcgaaattt caaagggaaa ttaaacgtct
     2401 atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttccacatc cgccacaaca
     2461 tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg
     2521 gccccgtgct gctgcccgac aaccactacc tgagcgtgca gaccaaactt tcgaaagacc
     2581 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2641 tcggcatgga cgagctgtac aagggcggta ccggagggat gatggtgaga aagggcgagg
     2701 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2761 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt
     2821 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2881 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2941 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggccact
     3001 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3061 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactcag
     3121 taatctccaa ggcaaagata cgcacagtga aaatgattgc cttggtactg gccgtcttcg
     3181 ccctctgctt tctgcctttc cacgtcacgc gcaccctcta ctactccttc cgatcacttg
     3241 acctcagctg ccacaccctc aacgccatca acatggcata taagatcacc cggccgctgg
     3301 ccagcgccaa cagttgtctt gacccggtac tctacttcct ggcagggcag agacttgtcc
     3361 gctttgcccg agatgccaag ccacccacgg agcctacccc cagcccacag gctcgtcgca
     3421 agctgggcct gcacaggcct aacagaactg tgaggaaaga tttgtcagtc agcagtgacg
     3481 actcaagacg gacagagtcc acaccagctg gaagtgagac taaggacatt cggctatagg
     3541 gatccactag ttctagagcg gccgcttcac tcctcaggtg caggctgcct atcagaaggt
     3601 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     3661 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     3721 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3781 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     3841 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     3901 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     3961 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     4021 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     4081 cttatgaaga tccctcgacc tggagctcca gcttttgttc cctttagtga gggttaattt
     4141 cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4981 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5461 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6061 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6181 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6241 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6301 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6421 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6601 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6721 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     6781 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6841 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6901 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggg
     6961 tacctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
